dc.contributor.author
Montacir, Othman
dc.contributor.author
Montacir, Houda
dc.contributor.author
Eravci, Murat
dc.contributor.author
Springer, Andreas
dc.contributor.author
Hinderlich, Stephan
dc.contributor.author
Mahboudi, Fereidoun
dc.contributor.author
Saadati, Amirhossein
dc.contributor.author
Parr, Maria Kristina
dc.date.accessioned
2018-06-08T11:07:17Z
dc.date.available
2018-03-22T13:55:33.225Z
dc.identifier.uri
https://refubium.fu-berlin.de/handle/fub188/21653
dc.identifier.uri
http://dx.doi.org/10.17169/refubium-24941
dc.description.abstract
Eptacog alfa (NovoSeven®) is a vitamin K-dependent recombinant Factor VIIa
produced by genetic engineering from baby hamster kidney (BHK) cells as a
single peptide chain of 406 residues. After activation, it consists of a light
chain (LC) of 152 amino and a heavy chain (HC) of 254 amino acids. Recombinant
FVIIa undergoes many post-translational modifications (PTMs). The first ten
glutamic acids of the N-terminal moiety are γ-carboxylated, Asn145 and Asn322
are N-glycosylated, and Ser52 and Ser60 are O-glycosylated. A head-to-head
biosimilarity study was conducted for the originator and the first biosimilar
AryoSeven™ to evaluate comparable bioengineering. Physicochemical properties
were analyzed based on mass spectrometry, including intact mass, PTMs and
higher-order structure. Both biotherapeutics exhibit a batch-to-batch
variability in their N-glycan profiles. N-Glycopeptide analysis with UHPLC-
QTOF-MSE confirmed N-glycosylation sites as well as two different
O-glycopeptide sites. Ser60 was found to be O-fucosylated and Ser52 had
O-glucose or O-glucose-(xylose)1,2 motifs as glycan variants. Ion mobility
spectrometry (TWIMS) and NMR spectroscopy data affirm close similarity of the
higher-order structure of both biologicals. Potency of the biodrugs was
analyzed by a coagulation assay demonstrating comparable bioactivity.
Consequently, careful process optimization led to a stable production process
of the biopharmaceuticals.
en
dc.format.extent
16 Seiten
dc.rights.uri
http://creativecommons.org/licenses/by/4.0/
dc.subject
biopharmaceutical
dc.subject
mass spectrometry
dc.subject
physicochemical characterization
dc.subject
coagulation assay
dc.subject.ddc
600 Technik, Medizin, angewandte Wissenschaften::610 Medizin und Gesundheit::615 Pharmakologie, Therapeutik
dc.subject.ddc
500 Naturwissenschaften und Mathematik::570 Biowissenschaften; Biologie::572 Biochemie
dc.title
Bioengineering of rFVIIa Biopharmaceutical and Structure Characterization for
Biosimilarity Assessment
dc.type
Wissenschaftlicher Artikel
dcterms.bibliographicCitation
Bioengineering 5 (2018), 7
dcterms.bibliographicCitation.doi
10.3390/bioengineering5010007
dcterms.bibliographicCitation.url
http://doi.org/10.3390/bioengineering5010007
refubium.affiliation
Biologie, Chemie, Pharmazie
de
refubium.affiliation.other
Institut für Chemie und Biochemie

refubium.mycore.fudocsId
FUDOCS_document_000000029385
refubium.note.author
Der Artikel wurde in einer Open-Access-Zeitschrift publiziert.
refubium.resourceType.isindependentpub
no
refubium.mycore.derivateId
FUDOCS_derivate_000000009573
dcterms.accessRights.openaire
open access